International Journal of Research in Medical Sciences
Chandra S et al. Int J Res Med Sci. 2017 Jan;5(1):46-49
www.msjonline.org
Original Research Article
pISSN 2320-6071 | eISSN 2320-6012
DOI: http://dx.doi.org/10.18203/2320-6012.ijrms20164520
Study of iron status indicators in different phases of menstrual
cycle in first year medical college females
Shipra Chandra1*, Narendra Gupta2, Sanjay Kumar Patel1
1
Department of Physiology, All India Institute of Medical Sciences, Delhi, India
Department of Physiology, Rohilkhand Medical College and Hospital, Bareily, Uttar Pradesh, India
2
Received: 29 November 2016
Accepted: 03 December 2016
*Correspondence:
Dr. Shipra Chandra,
E-mail: [email protected]
Copyright: © the author(s), publisher and licensee Medip Academy. This is an open-access article distributed under
the terms of the Creative Commons Attribution Non-Commercial License, which permits unrestricted non-commercial
use, distribution, and reproduction in any medium, provided the original work is properly cited.
ABSTRACT
Background: Iron requirements are increased in adolescent girls with growth and the onset of menarche and remain
high in women until menopause.
Methods: We conducted a study on first year medical college female students (age group 18-25 years) belonging to
socioeconomic classes 1 and 2 as per the Modified Kuppuswamy’s Scale to study the effect of the phases of
menstrual cycle on the iron status indicators (ISI) (Hb, Serum Iron, TIBC, MCV and TS%). Menstrual phases were
defined and blood samples were drawn from all the girls in all the 3 phases (menstrual, follicular and luteal) and Hb,
Serum iron and Total iron binding capacity were assessed on a semi-auto-analyzer and MCV was estimated using a
fully automated CBC machine. Transferrin Saturation was derived as Serum Iron / TIBC × 100.
Results: We found significant (p<0.01) difference in iron status indicators (ISI) that varied significantly in
accordance to the menstrual cycle, with values highest during the luteal phase and lowest during the menstrual phase.
Conclusions: The effect of hormonal fluctuations during the menstrual cycle plays a part in the variation of iron
status measures, which has to be considered while measuring them in a female during her reproductive years.
Keywords: Iron status indicators, Menstrual cycle
INTRODUCTION
Iron has several vital functions in the body. WHO
estimated that Iron Deficiency (ID) occurs in about 66–
80% of the world's population.ID has many negative
effects on health, including changes in immune function,
cognitive development, temperature regulation, energy
metabolism and work performance.1
ID and anemia is a worldwide problem that is highly
prevalent in developing regions of the world. In women,
ID has been associated with low dietary iron intake and
high levels of physical activity. The highest incidence of
anemia is reported in South Asia in women of
reproductive age and preschool children. It has serious
implications in terms of increased morbidity and
mortality rates in vulnerable groups, impaired growth and
cognitive abilities in children and reduced work capacity
and poor obstetric performance in adult women.2 In a
tropical country like India where infestations and
infections are a common occurrence in one’s life
combined with limited quality food intake, faulty cooking
and eating methods, the iron as well as the overall
nutritional status of females remains often compromised,
especially in the females who are more physically active.
In developing nations, iron deficiency (ID) and iron
deficiency anemia (IDA) occur most often in premenopausal women, mainly because of poor dietary
intake and the regular loss of blood during menstruation.3
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Chandra S et al. Int J Res Med Sci. 2017 Jan;5(1):46-49
The mean menstrual iron loss, averaged over the entire
menstrual cycle of 28 days, is about 0.56 mg/day. The
marked variation of menstrual losses is a great nutritional
problem.4 In boys, body iron stores may be increased
during and after their rapid growth period, whereas in
girls, iron stores remain low during the whole pubertal
period. The lower iron statuse in girls, besides growth,
are also because of the iron loss with menstruation.
Measurements of iron status in blood can be confounded
by non-nutritional factors. The effect of such
confounding factors must be considered when data are
used to evaluate the iron status of population groups. In
women of reproductive age, potential confounding
factors include the possible fluctuations in measures of
iron status associated with various phases of the
menstrual cycle.
Physiologic changes that occur during the menstrual
cycle, such as fluctuations in fluid volume, blood loss and
significant iron loss associated with it, may cause
variations in iron status indicators.5 So, while we conduct
a nation-wide survey to assess the nutritional status, it is
of foremost importance to assess, keeping in mind all the
factors that can alter present results, specially menstrual
cycle in females, girls at menarche, women right after
child birth, women suffering from irregular/heavy
menstrual cycle due to any gynecological disorder, post
traumatic and post major surgical patients, people
suffering from bleeding disorders. The present study was
designed to study the variations in iron-status measures in
young adult female students studying in first year of
medical college (18-25 years).
Aims and objectives
To study the iron status indicators in in young adult
female students studying in first year of medical college
(18-25 years) and to determine whether normal
physiological changes associated with menstrual cycle
affect the iron status indicators in the present study group.
METHODS
The study was conducted at Department of Physiology,
Moti Lal Nehru Medical College, Allahabad, Uttar
Pradesh, India between year 2011 to 2012. Approval
from the institute’s ethical committee and written
informed consent was taken from each girl participating
in the study.

Not on medication or hormonal supplements and of
same educational and SES.
Exclusion criteria







Non- compliant subjects
Irregular menstrual cycle
Any significant previous illness
Any bleeding disorder or hemoglobinopathy
History of major surgery, accident or blood
transfusion
Suffering from or suffered from any gynecological or
hormonal disorder
Family
history
of:
Any
blood
related
disorders/Menstrual irregularities/Infertility/Cancer
of female reproductive organs/Any gynecological
disorder.
Evaluation
After taking the informed consent, applying exclusion
and inclusion criteria the detailed history was taken
including name, age and the following:a) Dietary habits-veg./ non veg. with qualification and
quantification of the daily intake
b) Menstrual history with: Age at menarche & following
phase definition:



Menstrual phase: day 1-5, menstruating at the time of
survey
Follicular phase : 6-16 days after menstruation
Luteal phase: 17-30days after menstruation.
c) Detailed history regarding –Regularity/ Flow/
Debilitating pain/ Cyclical mastalgia/Last menstrual
period.
Complete general and systemic examination was carried
out on each subject as per the standard guidelines. The
following measurements were carried out on each
subject:
1. Anthropometry: Height, Weight and BMI
2. Iron status indicators:
a. Serum Iron: 11.6-31.7 µmol/lit in males & 9.0-30.4
µmol/lit in females.6
Inclusion criteria (n = 50)




b. Total Iron Binding Capacity6: 45 – 66 µmol/lit
Young adult unmarried female students of age 18-25
years studying in M.L.N. Medical College,
Allahabad.
Having normal menstrual cycle of 25-32 days
Living in same hostel
Eating from the same mess
c. Hemoglobin 7: 13.5-18 gm/dl in males and 11.5-16
gm/dl in females.
d. Transferrin Saturation6: 20 – 50 % in males and 1550% in females.
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Chandra S et al. Int J Res Med Sci. 2017 Jan;5(1):46-49
e. Mean Corpuscular Volume7: 74 - 95 µm3
Intra-cubital venous blood (5ml) was collected from the
subjects. The blood was allowed to stand for 60 minutes
in incubator at 37▫C. After that serum was obtained by
centrifugation. Fresh serum was used for SI and TIBC,
fresh whole blood for Hb and MCV determination.
Estimation of serum iron and TIBC was done by
ferrozine method (magnesium carbonate) obtaining the
absorbance using the semi-auto-analyzer by ARTOS
established in the Department of Physiology M.L.N.M.C
Allahabad.
Fe (III)
Acidic Medium
Fe (II)
Fe (II) + Ferrozine → Violet colored complex
Hemoglobin estimation was done by Cyan-methemoglobin method (Drabkin’s reagent), obtaining the
absorbance using the semi-auto-analyzer
Transferrin saturation = Serum Iron / TIBC × 100
Estimation of MCV was done by fully automated
complete blood count machine (Medonic M series)
installed in the Department of Pathology, M.L.N.
Medical College, Allahabad. Fresh un-clotted whole
blood was used for the MCV estimation.
Statistical analysis
The data obtained in the study group was expressed as the
mean ± standard deviation. Statistical analysis was done
with the help of statistical software SPSS 17.0. The p
value of < 0.05 was considered as statistically significant
RESULTS
Present results show difference in the iron status
indicators in the present study group. Also significant
correlation was found between the iron status indicators
during different phases of the menstrual cycle. Hb values
were significantly associated with the menstrual phase for
the study group.
Table 1: Mean and standard deviation of different
parameters in the study group.
Parameters
Age (years)
Height (cms)
Weight(kgs)
BMI (kg/m2)
SES
Study group (Mean ± S.D.)
21.32±1.95
157.60±6.09
51.94±5.65
20.90±1.42
1.58±0.72
Concentrations of Hb were lowest for girls in the
menstrual phase and highest for girls in the luteal phase.
Mean values of both SI and TS were also significantly
associated with phases of the menstrual cycle, their
values were lowest for girls in the menstrual phase and
highest for girls in the luteal phase. The patterns of TIBC
values were opposite to those observed for Hb, SI, and
TS.
Table 2: Mean and standard deviation of iron status
indicators during menstrual phase in the study group.
Parameters
Hemoglobin (gm/dlit)
Serum Iron (µmol/lit)
TIBC (µmol/lit)
TS%
MCV
Study Group (Mean ±S.D.)
11.85±1.01
16.37±1.46
62.37±1.96
26.28±3.11
89.51 ± 1.79
Measures of TIBC were highest for girls whose blood
was drawn in the menstrual phase and lowest during the
luteal phase. MCV values were different only during the
follicular phase, significantly lower than the values
observed during the menstrual phase. MCV values were
only slightly different during the luteal phase from the
menstrual phase (not significant). So, we found that the
values of iron status indicators vary according to the
menstrual cycle, with values highest during the luteal
phase and lowest during the menstrual phase. Also it is
seen that the females in the study group could be misdiagnosed as anemic if their menstrual phases were not
taken into account.
Table 3: Mean and standard deviation of iron status
indicators during follicular phase in the study group.
Parameters
Hemoglobin (gm/dlit)
Serum Iron (µmol/lit)
TIBC (µmol/lit)
TS%
MCV
Study Group (Mean ± S.D.)
12.14±1.01
17.01±1.49
61.71±2.00
27.63±3.22
88.37±1.80
Table 4: Mean and standard deviation of iron status
indicators during luteal phase in the study group.
Parameters
Hemoglobin (gm/dlit)
Serum Iron (µmol/lit)
TIBC (µmol/lit)
TS%
MCV
Study group (Mean ± S.D.)
12.42±1.01
17.67±1.53
61.01±2.01
29.01±3.37
89.23±1.78
DISCUSSION
Present findings suggest that females in the reproductive
age group (specially, during the active menstrual phase)
are more prone to lower ISI as compared to women not
actively menstruating and our findings are also confirmed
by the work of Clung MJP et al, Kim I et al, Bentely et al
and Elemo GN et al.3,5,8,9 The rhythms of ovarian
hormones during the course of the menstrual cycle
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Chandra S et al. Int J Res Med Sci. 2017 Jan;5(1):46-49
influence the secretion of hormones that control the
volume and content of the vascular space. Thus changes
in plasma volume and rise in plasma protein
concentrations are linked to the normal course of estrogen
and progesterone fluctuations during the menstrual
cycle.10 In present study we did not directly measure
changes in plasma volume, but our observations are in
accordance with the well-known hormonal changes of the
menstrual cycle and their known influence on hemoconcentration and dilution.
ACKNOWLEDGEMENTS
Hemo-concentration due to estrogen, mediated changes
in plasma volume or increased synthesis or secretion of
plasma proteins, might explain the mechanism for higher
iron-status values during the luteal phase. An increase in
plasma progesterone concentrations also occurs during
the luteal phase which has a natriuretic effect due to
aldosterone antagonism, resulting in plasma loss of
sodium and water.11
REFERENCES
Progesterone may also influence hemo-concentration
during the early to mid-luteal phase when its
concentrations peak. The fall in progesterone during the
luteal phase is associated with a sharp rise in aldosterone
activity observed few days before the onset of menses
manifesting as weight increase, edema & hemo-dilution.
Thus, the hormone-associated changes in plasma volume
and plasma protein concentration occurring during course
of the menstrual cycle offer a possible mechanism to
explain differences in the values of iron-status indicators
that we observed at different phases of the cycle.
CONCLUSION
From present observations we can conclude that the
effect of hormonal fluctuations during the menstrual
cycle has a part to play in the variation of iron status
measures, which has to be considered while measuring
them in a female during her reproductive years, as it may
alter the results depending on the phase of the menstrual
cycle.
Further modifications are possible in present study which
may clarify our understanding in this area even more. For
instance we could possibly study the levels of various
reproductive hormones and correlate with them. We
could also take more comparable parameters like serum
ferritin and transferrin receptor.
Authors would like to thank all subjects for participating
in the study who so willingly volunteered, without their
co-operation this study would not have been possible.
Funding: No funding sources
Conflict of interest: None declared
Ethical approval: The study was approved by the
Institutional Ethics Committee
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Cite this article as: Chandra S, Gupta N, Patel SK.
Study of iron status indicators in different phases of
menstrual cycle in first year medical college females.
Int J Res Med Sci 2017;5:46-9.
International Journal of Research in Medical Sciences | January 2017 | Vol 5 | Issue 1
Page 49